tbuckley/cabot-search huggingface dataset:
Abstract P2085: Whole Blood Microfluidics To Assess Direct Oral Anticoagulant (doac) Activity And Reversal In Neonatal Cardiac Patients
Authors: Thomas G Diacovo, Michael Decortin, Scott L Diamond
Journal: Circulation Research (2022)

Introduction: VTEs increase morbidity in neonates with congenital heart disease. Although heparin is the main therapy for prevention / treatment, DOACs are attractive alternatives due to ease of administration, no dependence on antithrombin and wider therapeutic window. Preclinical studies to evaluate their effects on clot formation and antidote reversal are hampered by a lack of low volume assays that permit multiple analyses. Hypothesis: A low volume microfluidic system can reproducibly measure activity and reversibility of DOAC-induced alterations in coagulation. Methods: A microfluidic device was used to study whole blood (WB) coagulation in response to co-immobilized tissue factor and collagen. Blood was drawn before and 24h after surgery. Platelet and fibrin deposition were monitored by fluorescence microscopy (Apixaban ± andexanet). Results: 34 neonates were enrolled. Most participants had double ventricle physiology, required bypass and blood products. There was no difference (P&gt;0.05) in Hb / Hct, platelet count, or fibrinogen levels pre- vs post-surgery. Time to clot-induced channel occlusion and platelet / fibrin deposition were similar in post-operative vs adult participants (7.6±3.1 min vs 7.9±1.5 min; Figure 1A), the former prolonged (14.2±1.9 min) and the latter reduced in pre-operative patients. Although Apixaban reduced clot formation (Figure 1B), it prevented channel occlusion in most post-operative neonates vs adults (60% vs 20%). Andexanet reversed its effects, restoring coagulation to non-treatment levels. Conclusions: WB microfluidics demonstrated that post-operative neonates are hypercoagulable compared to the pre-operative state and are responsive to apixaban, the effects of which can be reversed with Andexanet.
Pancoast syndrome: an unusual presentation of adenoid cystic carcinoma
Authors: MQ Hatton, MB Allen, NJ Cooke
Journal: European Respiratory Journal (1993)

We report on a patient with primary pulmonary adenoid cystic carcinoma presenting with Pancoast syndrome. Pancoast syndrome has not previously been described with this tumour. Other unusual features of this case include the peripheral origin and mediastinal involvement, with lack of proximal endobronchial spread.
Abstract P3062: T-cell Myd88 Regulates Cardiac Fibrosis In Heart Failure By Modulating T-cell Survival And Pro-inflammatory Signaling
Authors: Abraham L Bayer, Sasha Smolgovsky, Njabulo Ngwenyama, Mark Aronovitz, Kuljeet kaur, Pilar Alcaide
Journal: Circulation Research (2022)

Background: Despite the association of heart failure (HF) with cardiac and systemic inflammation, anti-inflammatory therapies have shown limited success. Cardiac inflammation is characterized by the presence of damage associated molecular patterns (DAMPs), myeloid and T-helper cell infiltration, and the activation of cardiac fibroblasts (CFB). The adaptor “Myeloid differentiation response 88” (MyD88) is central to DAMP sensing in myeloid cells, but its role in T-cell function and cardiac inflammation is largely unknown. We hypothesized that T-cell signaling through MyD88 modulates cardiac inflammation and fibrosis in HF. Methods and Results: We reconstituted T-cell deficient ( Tcra -/- ) mice, normally protected from cardiac fibrosis, with WT or Myd88 -/- Type 1 helper T-cells (Th1) in the onset of transaortic constriction (TAC). We found mice given Myd88 -/- Th1 cells exhibited significantly higher levels of cardiac T-cell infiltration, increased cardiac fibrosis, and lower fractional shortening than mice given WT Th1 cells. T-cell specific MyD88 deficient mice ( Myd88 fl/fl CD4 Cre ) also developed more fibrosis and increased cardiac T-cells compared to Cre- littermate controls in response to TAC. RNA-seq analysis of WT and Myd88 -/- Th1 cells revealed enrichment of pro-inflammatory signaling and pro-survival genes in Myd88 -/- Th1 cells, which we validated by immunoblotting for phospho-ERK, NF-kB, and AKT. In contrast, IFNγ, the Th1 signature cytokine, was similar by qPCR and flow cytometry in WT and Myd88 -/- cells. Moreover, we discovered that Myd88 -/- Th1 cells have increased survival in vitro, by monitoring propidium iodide incorporation in real time, and in vivo by adoptive transfer of equal numbers of WT and Myd88 -/- cells together into Tcra -/- mice and quantifying both cell types in lymphoid organs over time. Lastly, more Myd88 -/- Th1 cells adhered to immobilized VCAM-1 and CFB in vitro compared to WT, resulting in enhanced CFB transformation. Conclusion: T-cell MyD88 limits cardiac fibrosis in TAC through modulation of T-cell inflammatory signaling and cell survival. Disruption of this pathway in T-cells results in enhanced transformation of fibroblasts and worsened systolic function.
Abstract P1035: Co-culturing Human Induced Pluripotent Stem Cell-derived Cardiomyocytes With Mesenchymal Stem Cells Promotes Their Maturation And Enhances Cardioprotection Under Hypoxic Stress
Authors: Divya Sridharan, Uzair Ahmed, Mahmood Khan
Journal: Circulation Research (2022)

Introduction: Co-culturing human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (hiCMs) with other stem/progenitor cells like bone-marrow mesenchymal stem cells (BM-MSCs) and endothelial progenitors has been shown to improve their survival post transplantation into ischemic hearts. Co-culturing with other cardiac cells has also shown to improve the maturation of hiCMs in vitro. However, the potential of co-culturing hiPSC-derived MSCs (hiMSCs) for hiCM maturation and cardioprotection has not been explored. Hypothesis: We hypothesize that co-culturing of hiMSCs with hiCMs significantly alters the cells’ proteome, improves hiCM maturation, and enhances cardioprotection. Results: hiCMs were co-cultured with hiMSCs for one week at a ratio of 3:1. To evaluate the effect of hiMSCs on hiCM maturation in the co-culture, changes in the hiCM function and gene expression were assessed via multi electrode array (MEA) analysis and qPCR, respectively. MEA analysis showed an increase in ADP50 and ADP90 in addition to improved response to cardiac drugs in hiCMs co-cultured with hiMSCs as compared to hiCM-only cultures, indicating improved maturation of hiCMs in the co-culture. Furthermore, gene expression analysis showed an increased expression of mature CM-associated genes (MYH7, TNNT2). When subjected to hypoxic stress (1% O 2 ), we also observed a significant decrease in the number of TUNEL positive cells in the co-cultures as compared to hiCM-only culture. Assessment of hiCM functionality via MEA analysis showed an improved cardioprotection in co-cultures as compared to the single cell-type cultures. Furthermore, proteomic analysis demonstrated increased expression of cardioprotective growth factors (GFs): HGF, VEGFA and IGFBP-1 in the conditioned medium of the co-cultured cells versus single cell-type cultures. Conclusion: Overall, our results established that co-culturing hiCMs with hiMSCs improved cardioprotection via increased the expression of cardioprotective growth factors and enhanced the maturation of hiCMs in vitro.
Abstract P3128: Obesogenic Diet Promotes Endothelial-to-Mesenchymal Transition In Adipose Tissue
Authors: Nicholas W Chavkin, Karen Hirschi, Noyan GOKCE, Kenneth Walsh
Journal: Circulation Research (2022)

Vascular dysfunction and chronic inflammation are characteristics of obesity-induced adipose tissue dysfunction. Proinflammatory cytokines can drive Endothelial-to-Mesenchymal Transition (EndoMT), where endothelial cells transition into mesenchymal-like interstitial cells that leave the vascular wall and promote tissue inflammation and fibrosis. In this study, we aimed to determine whether obesogenic diet can promote EndoMT in adipose tissue. EndoMT was assessed in 1) transgenic endothelial cell lineage-tracing mice subjected to High-Fat/High-Sucrose (HF/HS) diet, 2) available single cell RNA sequencing datasets from human obese adipose tissue, and 3) cell culture models using primary human adipose-derived endothelial cells. In lineage-tracing mice, obesogenic diet induced EndoMT in a time-dependent manner at 13, 26, and 52 weeks on HF/HS diet in both subcutaneous and visceral adipose tissue. Interestingly, rate of EndoMT was greater in visceral compared to subcutaneous adipose tissue. After 52-weeks on HF/HS diet, EndoMT cells in visceral adipose tissue upregulate inflammatory response and TGF-Beta signaling transcriptional pathways compared to endothelial cells in the same adipose tissue depot. Initiation of EndoMT was also found in single cell RNA sequencing datasets of stromal cells from human obese adipose tissue, indicated by endothelial cells in these datasets containing sub-clusters with reduced Pecam1 and increased Acta2 expression. Finally, the capacity for EndoMT in primary human adipose-derived endothelial cells was validated by bulk RNA sequencing after treatment with proinflammatory cytokines (TNF-Alpha, IFN-Gamma, TGF-Beta1), confirming decreased expression of endothelial cell genes and increased expression of genes associated with EndoMT, inflammatory response, and TGF-Beta signaling. Together, these clinical and experimental findings indicate that chronic obesity can promote EndoMT in adipose endothelial cells.
Abstract P2049: Myofibroblast Depletion Of Yap And Wwtr1 Improves Cardiac Function After Myocardial Infarction Via Ccn3 Suppression
Authors: Michael A Flinn, Santiago Alvarez, Makenna C Knas, Victor Alencar, Samantha Paddock, Xiaoxu Zhou, Tyler Buddell, Ayana Jamal, Pengyuan Liu, Jenny Drnevich, Michaela Patterson, Brian Link, Caitlin Omeara
Journal: Circulation Research (2022)

The Hippo-Yap pathway is an area of interest in cardiac fibroblasts, as inhibition Lats1/2 promotes substantial fibrosis and Yap/Wwtr1 facilitate maturation of fibroblasts to myofibroblasts. While the Hippo-Yap pathway has been characterized in resting and activated fibroblasts, we take the novel approach of investigating its role in myofibroblasts. We assess depletion of Yap alone ( Yap fl/fl ;Postn MCM ) or Yap and Wwtr1 ( Yap fl/fl ;Wwtr1 fl/+ ;Postn MCM ) in myofibroblasts immediately following myocardial infarction and focus on identifying and validating downstream factors mediating pathological remodeling in mice. Following injury, depletion of Yap alone had no effect on heart function, but depletion of Yap and Wwtr1 resulted in improved ejection fraction and fractional shortening at 60 days post injury. Yap fl/fl , Wwtr1 fl/+ ;Postn MCM hearts also displayed smaller scars, reduced interstitial fibrosis, and more denatured collagen. Single cell RNA sequencing of interstitial cells 7 days post injury showed suppression of pro-fibrotic and pro-inflammatory genes and suppression of a matrifibrocyte phenotype in fibroblasts derived from the Yap fl/fl , Wwtr1 fl/+ ;Postn MCM heart. Analysis of in vivo and in vitro transcriptomics revealed depletion of Yap/Wwtr1 resulted in dramatically decreased fibroblast expression of the matricellular protein Ccn3. Administration of recombinant CCN3 to mice following injury aggravated cardiac function and scarring over the course of 28 days. Thus, we show Yap/Wwtr1 depletion in myofibroblasts attenuates fibrosis and significantly improves cardiac outcomes after injury and identify Ccn3 as a novel factor that contributes to adverse cardiac function. Given therapeutic strategies would likely be implemented after myocardial infarction when myofibroblasts are already activated, our work suggests targeting Yap and Wwtr1, or Ccn3 directly as an approach for suppressing pathological remodeling.
Abstract P1132: Cardiac Tissue Chip Model Of Arteriovenous-fistula-associated Hemodynamics Elicits Fibrotic Changes Seen In Mouse Model
Authors: Caleb Graham, Tatyana Isayeva-Waldrop, William Gard, Kevin Ingle, Timmy Lee, Palaniappan Sethu
Journal: Circulation Research (2022)

Introduction: Cardiovascular disease is the primary cause of death amongst end-stage renal disease (ESRD) patients. Though arteriovenous fistulas (AVFs) are the access of choice for hemodialysis, joining a high-pressure artery and low-pressure vein leads to cardiac volume overload acutely. Debate exists as to the cardiac ramifications of AVFs, due in part to the difficulty of recruiting ESRD patients and controls for prospective studies. Animal models have been essential in AVF studies, but their usefulness is limited by interspecies differences in physiology and other factors. Thus, we have developed a three-dimensional (3D) cardiac tissue chip with tunable pressure and stretch to model the acute hemodynamic changes associated with AVF creation. Hypothesis: In this study, we aimed to replicate the hemodynamics of murine AVF models in vitro and hypothesized that if 3D cardiac constructs were subjected to “volume overload”, they would display fibrosis and key gene expression changes seen in AVF mice. Methods: Mice underwent either AVF or sham procedure and were monitored for 28 d. Cardiac tissue constructs composed of h9c2 rat cardiac myoblasts and normal adult human dermal fibroblasts in hydrogel were seeded into devices and exposed for 96 h to 100 mmHg/10 mmHg pressure (0.4 s/0.6 s) at 1 Hz. "Systolic" pressure was generated via pneumatic pump while a hydrostatic pressure head gave rise to "diastolic" pressure. The controls were exposed to “normal” stretch while the experimental group was exposed to “volume overload”. Results and Conclusions: The in vitro model yielded similar fibrosis-related gene expression profiles (see figure) and patterns of fibrosis as the murine model.
Abstract P1048: Dual Myristic Acid And Trans Activator Of Transcription Conjugated Peptide Facilitates Delivery Of Protein Kinase C Beta II Peptide Inhibitor To Attenuate Myocardial Ischemia/Reperfusion Injury
Authors: Logan Clair, James Ramsarran, Taurai Augustin, Cameron Stinson, Tameka Dean, Qian CHEN, Robert Barsotti, Lindon Young
Journal: Circulation Research (2022)

Peptides conjugated to myristic acid (myr) or trans activator of transcription (tat) have long been used to increase cell permeability via simple diffusion and endocytosis, respectively. Previously myr-PKCβII inhibitor (myr-PKCβII-) exerted cardioprotective effects in myocardial ischemia/reperfusion (I/R) and inhibited superoxide (SO) release from rat leukocytes by 40% (20 μM). By contrast, dual conjugated myr-tat-PKCβII inhibitor (myr-tat-PKCβII-) inhibited SO release by 90% at the same concentration. The purpose of this study was to determine the potency of myr-tat-PKCβII- compared to myr-PKCβII- in mitigating infarct size and restoring cardiac function in an ex-vivo rat model of myocardial I/R injury. Isolated hearts from male Sprague-Dawley rats (~300g) were subjected to global I(30-min)/R(50-min) . Left ventricular cardiac function was recorded using a pressure transducer. Treatments were infused during the first 5 min of R. After R, the hearts were sectioned (2 mm) from base to apex and stained with 1% triphenyltetrazolium chloride. Infarcted tissue was excised to determine infarct size (i.e. infarct tissue weight/ total tissue weight). Data were analyzed using ANOVA Fisher’s LSD analysis. Compared to untreated controls (23.2±3.1%, n=17), myr-tat-PKCβII- exerted a significant and similar decrease in infarct size that was observed from 100 nM (9.7±0.59%, n=3, p&lt;0.05), 1 nM (10.0±2.3%, n=5, p&lt;0.05), to 100 pM (12.6±2.3%, n=5, p&lt;0.05); but not at 10 nM (13.8±3.6%, n=5). Cardiac function for all treatment groups did not significantly improve from control. However, at 100pM (1070±170 mmHg/s), +dP/dt max was significantly improved compared to all other treatment groups (p&lt;0.05). The addition of myr to tat-conjugated peptides improved the cardioprotective effects ~200-fold compared to myr-PKCβII- and ~2000-fold compared to native peptide presumably by enhanced intracellular delivery of cargo. The ~50% reduction in infarct size suggests that myr-tat-PKCβII- is cardioprotective in I/R. Future studies will test our most effective dose determined in ex vivo hearts to restore post-reperfusion cardiac function combined with significant cardiac salvaging effect (reduced infarct size) in a porcine myocardial I/R model.
Abstract P1069: Integrin Linked Kinase Controls Human Valve Endotelial Cells Pro-osteogenic Activation In A TFG-b/NO Dependent Manner
Authors: Sandra Sanchez, Mercedes Castro, Alberto Cook Calvete, María Delgado Marín, Ignacio Hernandez Navarro, Jose Lopez-Menendez, Carlos Zaragoza, Marta Saura
Journal: Circulation Research (2022)

Calcific aortic valve disease (CAVD) is expected in the aging population. CAVD physiopathology arises from endothelial dysfunction leading to lipid accumulation, inflammation, and osteogenic transformation. Many studies suggest that valve endothelial cells (VECs) undergoing EndMT may differentiate into osteoblastic interstitial cells. We previously showed that Integrin linked kinase (ILK) regulates vasomotor function by preventing endothelial nitric oxide synthase uncoupling. ILK expression in vascular endothelium plays an essential cardioprotective role. Moreover, we have found a significant negative correlation between human valve protein levels of ILK and the osteogenic marker Runx2 and BMP2 in 70 patients with calcific aortic stenosis, a higher decrease at the endothelial level. We hypothesized that Endothelial ILK plays a protective role in human valve endothelial cells osteogenic transition. Isolation of human valve endothelial cells from the aortic (aortic hVECs) and ventricular sides (ventricular hVECs) show a specific ILK decrease in the aortic hVECs from calcific compared to non-calcified valve (p&lt;0.05). Silencing ILK expression in hVECs decreased NO production and increased ROS (Reactive Oxygen Species) production as early as 72 hours. After five days of silencing ILK, hVEC express myofibroblast markers SM22a, α-SMA, MMP12, MMP9, and SNAI 1 while losing the endothelial markers (CD31, and vWF (p&lt;0.01). ILK silenced hVECs (siILK-hVECs) express the osteogenic marker RunX2 indicating osteogenic differentiation and developed calcified nodules. Cell treatment with Noggin, a BMP2 antagonist, did not reverse the increase of Runx2, thus excluding the implication of the BMP2/RUNX2 axis. However, silk-hVECs showed an increase in phosphorylated-SMAD 2, suggesting a TFG-β -dependent mechanism. Treatment of siILK-hVECs with the NO donor DETA-NO decreased Smad 2 activation and RunX2. Moreover, DETA-NO treated siILK hVECs cultured in pro-osteogenic media did not develop calcification. Collectively, our results point to a crucial role of ILK in keeping human valve endothelial cell phenotype preventing valve calcification in a Nitric Oxide-TFGβ1 dependent manner. Financed by the Regional European fund “a way to achieve Europe.” JCCM SBPLY/19/180501/000055S and ISCIII PI20/00930
Abstract P3045: The Role Of Mitochondrial Dysfunction In Arrhythmogenesis
Authors: Akash D Chakraborty, Ying Ann Chiao, Chi Fung Lee
Journal: Circulation Research (2022)

Cardiac arrhythmia is triggered by deregulated ion channels and abnormal electrical activity, and it increases susceptibility of patients to sudden death. The regulations of ion movement in and out of the myocytes or subcellular compartments are highly energy-dependent. Although impaired energetics has been implicated in arrhythmia, the causal roles and intricate details of how mitochondrial dysfunction contributes to arrhythmogenesis are far from established. We employed a mouse model with cardiac-specific mitochondrial dysfunction by deleting Ndufs4, a protein of mitochondrial complex I (Ndufs4-cKO). Complex I deficiency in Ndufs4-cKO mice lowered cardiac NAD/NADH ratio but did not affect cardiac function and geometry at baseline. Control and Ndufs4-cKO mice showed normal sinus rhythm at baseline. Ndufs4-cKO mice had 3-fold increases non-sinus arrhythmic events upon an arrhythmogenic stress test. 50% of Ndufs4-cKO mice succumbed to sudden cardiac death with displays of pre-ventricular contractions and ventricular fibrillation. We collected cardiac tissues at baseline and post stress. We observed increased ryanodine receptor, RyR2-S2814 phosphorylation, reduced phospholamban PLN S16 phosphorylation and decreased calsequestrin 2 expression in Ndufs4-cKO hearts post stress, but not at baseline. Isolated Ndfus4-cKO cardiomyocytes after stress test show a trend towards prolonged time to peak shortening and time to peak Ca 2+ transient. In addition, elevating cardiac NAD levels lowered arrhythmic events of Ndufs4-cKO mice after stress test. These results revealed direct evidence that mitochondrial dysfunction increases susceptibility of hearts to arrhythmia by regulating NAD redox balance and SR calcium homeostasis.Ndufs4-cKO mice with chronic high fat diet (HFD) had accelerated declines in systolic and diastolic function and increased hypertrophy, compared to control mice. WT mice with HFD showed diastolic dysfunction and increased arrhythmic events after stress test, while diastolic dysfunction was reversed by elevating cardiac NAD levels. Our data suggest an arrhythmogenic mechanism linking mitochondrial dysfunction and NAD redox imbalance with SR calcium homeostasis, which may explain HFD-induced arrhythmogenesis.
Abstract P1044: A Drug-Like Activity Of Polydopamine Nanoparticles Mediates Cardiomyocyte Bio-Modulation And Maturation
Authors: Syed Baseeruddin Alvi, Divya Sridharan, Mahmoud Shalaan, muhamad mergaye, Mahmood Khan
Journal: Circulation Research (2022)

Introduction: Bio-active nanomaterials are being extensively explored for various biomedical and tissue engineering applications. Polydopamine nanoparticles (PDA-NPs) is a self-assemble polymer derived from dopamine which has demonstrated anti-inflammatory potential by modulating immune cells. The use of PDA-NPs as bio-active nanomaterial for cardiac tissue repair/regeneration warrants further exploration. Hypothesis: We hypothesize that the polydopamine nanoparticles could modulate cardiomyocyte activity due to the presence of catechol moieties and may alter the gene expression and promote maturation of cardiomyocyte. Results: The PDA-NPs were fabricated by oxidative polymerization of dopamine monomers which resulted in formation of spherical nanoparticles. These nanoparticles were in the range of 100- 200 nm as evident from SEM and TEM imaging. The FTIR analysis of PDA-NPs revealed the characteristic peaks confirming the presence of quinone and catechol groups. These functional groups play important role in surface adhesion, thus imparting adhesive properties to PDA-NPs. The PDA-NPs were exposed to human iPSC-derived cardiomyocytes (HiPSC-CMs) and their real-time activity was monitored by using MEA. An instantaneous change in the beat period of cardiomyocytes was observed in a dose dependent manner. Moreover, a significant shortening in field potential duration was also seen, these finding suggest that the PDA-NPs stimulates the cardiomyocytes through catechol groups thereby exhibiting strong adrenergic effect. These finding demonstrates that PDA-NPs could significantly modulate the contractility of cardiomyocytes and could be used for enhancing cardiac contractility. Furthermore, prolonged incubation of PDA-NPs showed accumulation in the perinuclear region as revealed by fluorescence imaging. The influence of intracellular accumulation of PDA-NPs on gene expression was assessed by qRT-PCR. A significant increase in MYL2/MYL7 ratio, GJA5 and TNNI3 was noted, demonstrating the maturation of cardiomyocytes. Conclusion: Overall, the treatment of hiPSC-CMs cardiomyocytes with PDA-NPs shows dual response, primarily by modulating the contractility in a dose dependent manner and secondary by promoting maturation.